Disclosure of Invention
In view of the above, the invention provides a light environment regulation method for delaying the growth of potato tissue culture seedlings. The method provides a scientific and reasonable light source providing scheme for prolonging the subculture period of a plant tissue culture factory and storing potato tissue culture seedlings (test tube seedlings), improves the electric energy utilization rate and stabilizes the quality of the potato tissue culture seedlings (test tube seedlings) while prolonging the subculture period of the potato tissue culture seedlings (test tube seedlings) in the full-artificial light plant tissue culture factory.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a light environment regulation and control method for delaying the growth of potato tissue culture seedlings, which is characterized in that the potato tissue culture seedlings are subcultured and cultured under the condition of a full artificial light source, and the full artificial light source condition is as follows according to the integral percentage of a photophysical spectrum: 0-50% of red light, 30-100% of blue light, 0-45% of green light and 0-20% of full visible light continuous spectrum, wherein the sum of the percentages is 100%;
the peak wavelength of the red light is 610-660 nm, the peak wavelength of the blue light is 430-480 nm, and the peak wavelength of the green light is 540-560 nm.
The method utilizes the targeted influence of light environments with different wavelengths on the synthesis and distribution of plant self-hormones, proteins and the like, and realizes the growth and development process of the potato tissue culture seedlings (test-tube seedlings) under the luminous environment by adjusting the proportion of light sources in each wave band in an artificial light source in the culture process of the potato tissue culture seedlings (test-tube seedlings). On the premise of following the growth rule of plants, the invention reduces the using amount of hormone, improves the energy utilization efficiency and reduces the production cost.
The LED artificial light source can emit wave bands which are required by plants and have the effective utilization rate of 80-90%, and can realize independent control of different light qualities and luminous intensities, thereby meeting the special requirements of facility agriculture on light source equipment. In addition, the LED is also suitable for being applied to artificial regulation and control plant cultivation environments with higher planting density, such as plant tissue culture, facility agriculture, industrial seedling culture, space ecological life-preserving systems and the like.
Preferably, the total artificial light source conditions are as follows according to the integral percentage of the light mass spectrum: 10-45% of red light, 30-80% of blue light, 0-40% of green light and 0-15% of full visible light continuous spectrum, and the sum of the percentages is 100%.
Preferably, the total artificial light source conditions, in terms of light mass spectral integral percentage, are: 20 to 45 percent of red light, 30 to 60 percent of blue light, 10 to 40 percent of green light and 0 to 15 percent of full visible light continuous spectrum, wherein the sum of the percentages is 100 percent.
In the present invention, the cultivation is carried out in two stages, including a first subculture stage and a second subculture stage;
the first group of culture stages is 0-48 h after subculture inoculation;
the second group culture stage is 48h after subculture.
Preferably, the total artificial light source of the first group culture stage is as follows: 20 to 45 percent of red light, 30 to 60 percent of blue light, 10 to 40 percent of green light and 0 to 15 percent of full visible light continuous spectrum, wherein the sum of the percentages is 100 percent;
the intensity of illumination received by the top end of the tissue culture bottle in the first group of culture stages is 20-50 mu mol/(m)2S); preferably, the intensity of the illumination received by the top end of the tissue culture bottle in the first tissue culture stage is 20-40 mu mol/(m)2·s)。
The illumination time of the first group culture stage is 12-14 h/24 h.
In the invention, the total artificial light source is direct light and scattered light.
Preferably, the total artificial light source of the first set of cultivation stages is: 30 percent of red light, 30 percent of blue light, 30 percent of green light and 10 percent of full visible light continuous spectrum, and the sum of the percentages is 100 percent;
the illumination intensity received by the top end of the tissue culture bottle in the first group of culture stages is 30 mu mol/(m)2·s);
The illumination time of the first group culture stage is 14h/24 h.
Preferably, the total artificial light source of the second culturing stage is as follows: 30-45% of red light, 30-45% of blue light, 10-35% of green light and 0-10% of full visible light continuous spectrum, wherein the sum of the percentages is 100%;
the illumination intensity received by the top end of the tissue culture bottle in the second tissue culture stage is 50-120 mu mol/(m)2S); preferably, the illumination intensity received by the top end of the tissue culture bottle in the second tissue culture stage is 55-120 mu mol/(m)2·s);
The illumination time of the second group culture stage is 14-16 h/24 h.
Preferably, the total artificial light source of the second culturing stage is: 30 percent of red light, 40 percent of blue light and 30 percent of green light, wherein the sum of the percentages is 100 percent;
the illumination intensity received by the top end of the tissue culture bottle in the second tissue culture stage is 110 mu mol/(m)2·s);
The illumination time of the second group culture stage is 16h/24 h.
Preferably, the culture temperature is 15-25 ℃, and the air humidity is 30-60%.
Preferably, the temperature of the culture is 19-25 ℃.
In the specific embodiment provided by the invention, the temperature for culturing is 19-22 ℃ or 20-25 ℃.
Preferably, the total artificial light source is a strip light source, the distance between two adjacent strip light sources is 0.10-0.46 m, the height of the strip light source from the top end of the tissue culture bottle is 0.10-0.20 m, and the light-emitting angle of the strip light source is 90-120 degrees.
The culture method provided by the invention gives different artificial light sources in different growth periods of the potato tissue culture seedlings (test tube seedlings), and the light energy obtained in the growth process of the potato tissue culture seedlings (test tube seedlings) is provided by the artificial light sources, and does not relate to direct solar light and scattered light, so that the culture method can be applied to the construction of plant tissue culture factories which mainly use the artificial light sources.
In the invention, when the potato tissue culture seedling (test-tube seedling) does not need to be stored continuously, the subculture tissue culture can be carried out on the potato tissue culture seedling (test-tube seedling), and the potato tissue culture seedling (test-tube seedling) can be placed under the condition of light environment suitable for the growth of the potato tissue culture seedling (test-tube seedling) to continue to grow.
The invention provides a light environment regulation and control method for delaying the growth of potato tissue culture seedlings. The light environment regulation and control method comprises the following steps: subculturing and inoculating the potato tissue culture seedlings, and culturing under the condition of a full artificial light source, wherein the full artificial light source condition is as follows according to the integral percentage of a light mass spectrum: 0-50% of red light, 30-100% of blue light, 0-45% of green light and 0-20% of full visible light continuous spectrum, wherein the sum of the percentages is 100%; the peak wavelength of the red light is 610-660 nm, the peak wavelength of the blue light is 430-480 nm, and the peak wavelength of the green light is 540-560 nm. Compared with the prior art, the invention has the following beneficial effects:
(1) the invention discloses a light environment regulating and controlling method for delaying growth of potato tissue culture seedlings (test-tube seedlings), which is used for setting light environment parameters for prolonging the subculture period of potato tissue culture seedlings (test-tube seedlings) in a total artificial light plant tissue culture factory, and relates to a light environment parameter for prolonging the subculture period and storing the growth of potato tissue culture seedlings (test-tube seedlings) in the total artificial light plant tissue culture factory. According to the invention, on the premise of following the growth rule of the plant, the specific influence of light environments with different wavelengths on the synthesis and distribution of plant hormones, proteins and the like is utilized, the growth development process of the potato tissue culture seedling (test-tube seedling) is prolonged by adjusting the light environment through adjusting the proportion of light sources of all wave bands in an artificial light source in the culture process of the potato tissue culture seedling (test-tube seedling), the use amount of hormones is reduced, the energy utilization efficiency is improved, and the production cost is reduced.
The invention does not relate to the addition of any exogenous additive (hormone) or microorganism, covers a group of culture physiological growth periods that potato tissue culture seedlings (test-tube seedlings) grow to bottle-formed seedlings after subculture inoculation, utilizes the influence of light on the growth and differentiation of plant callus to regulate and control the production and distribution of the growth hormone of the potato tissue culture seedlings (test-tube seedlings), prolongs the subculture period of the potato tissue culture seedlings (test-tube seedlings) in a full-artificial light plant tissue culture factory, reduces the intake of exogenous additive and microorganism, and reduces the pollution rate of the potato tissue culture seedlings (test-tube seedlings). After the light environment condition is changed, the potato tissue culture seedling (test-tube seedling) is easy to recover growth in subculture, and is suitable for the production purposes of prolonging the subculture period, preserving the potato tissue culture seedling (test-tube seedling) and the like.
(2) The light environment regulation and control method for delaying the growth of the potato tissue culture seedlings (test-tube seedlings) has wide coverage range. According to the facility conditions of the current mainstream plant tissue culture factory, and in combination with the change conditions of the external temperature and humidity environment, a light environment regulation and control method for delaying the growth of the potato tissue culture seedlings (test-tube seedlings) is worked out.
(3) The illumination scheme is highly targeted. The method aims at prolonging the subculture period and meeting the requirement of the potato tissue culture seedling (test-tube seedling) on the light environment, and sets a proper illumination scheme for storing the tissue culture seedling (test-tube seedling).
Detailed Description
The invention discloses a luminous environment regulation and control method for delaying the growth of potato tissue culture seedlings, and a person skilled in the art can appropriately improve process parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The test materials used in the present invention are all available on the market.
The invention is further illustrated by the following examples:
example 1
The embodiment provides a light environment regulation and control method for delaying growth of potato tissue culture seedlings (test-tube seedlings), which is used in a plant tissue culture factory with an artificial light source as a main light source, namely, light energy obtained in the growth process of the tissue culture seedlings (test-tube seedlings) is provided by the artificial light source, and direct light and scattered light of the sun are not involved; when potato tissue culture seedlings (test-tube seedlings) are planted in the plant tissue culture factory, the artificial light source specifically comprises:
after 0-48 h (the first group culture stage) and 48h (the second group culture stage) of the potato tissue culture seedlings (test-tube seedlings) after subculture inoculation, the artificial light source comprises the following components in percentage by mass according to the light-mass spectrum integral: 45 percent of red light, 30 percent of blue light, 15 percent of green light and 10 percent of full visible light continuous spectrum, the peak wavelength of the red light is 610-660 nm, the peak wavelength of the blue light is 430-480 nm, the peak wavelength of the green light is 540-560 nm, and the total illumination intensity of an artificial light source received by the top end of the tissue culture bottle in the first group of culture stages is 20 mu mol/(m2S) direct and scattered light, the total illumination intensity of the artificial light source received by the top of the tissue culture bottle in the second tissue culture stage being 55 μmol/(m)2S) direct light and scattered light. The accumulated illumination time of the artificial light source of the plant tissue culture factory in all tissue culture stages within 24 hours is 14 hours, the growth environment temperature of tissue culture seedlings (test tube seedlings) is 20-25 ℃, and the air humidity is 30-60%.
In a plant tissue culture factory, strip-shaped light sources are arranged on each layer of tissue culture rack, the distance between two adjacent artificial strip-shaped light sources is l, and the calculation formula of l is shown as (I):
in the formula (I), alpha is a light-emitting angle and the unit is DEG;
h is the height from the light source to the top end of the tissue culture bottle, and the unit is m.
In this example, α is 90 °; h is 0.11 m; l is 0.16 m.
In the plant tissue culture factory using the artificial light source as the main light source in the embodiment 1, that is, all light energy obtained in the growth process of the tissue culture seedling (test-tube seedling) is provided by the artificial light source, and does not relate to direct solar light and scattered light), the potato tissue culture seedling is cultured, and the height of the tissue culture seedling above the culture medium 24-26 days after inoculation can reach 6-7 cm.
Example 2
The embodiment provides a light environment regulation and control method for delaying growth of potato tissue culture seedlings (test-tube seedlings), which is used in a plant tissue culture factory with an artificial light source as a main light source, namely, light energy obtained in the growth process of the tissue culture seedlings (test-tube seedlings) is provided by the artificial light source, and direct light and scattered light of the sun are not involved; when potato tissue culture seedlings (test-tube seedlings) are planted in the plant tissue culture factory, the artificial light source specifically comprises:
after 0-48 h (the first group culture stage) and 48h (the second group culture stage) of the potato tissue culture seedlings (test-tube seedlings) after subculture inoculation, the artificial light source comprises the following components in percentage by mass according to the light-mass spectrum integral: continuous spectrum of 40% red light, 40% blue light, 10% green light and 10% full visible light, peak wavelength of red light is 610-660 nm, peak wavelength of blue light is 430-480 nm, peak wavelength of green light is 540-560 nm, total illumination intensity of artificial light source received by the top end of the tissue culture bottle in the first group of culture stages is 35 mu mol/(m)2S) direct and scattered light, the total illumination intensity of the artificial light source received by the top of the tissue culture bottle in the second tissue culture stage being 100 [ mu ] mol/(m)2S), the accumulated illumination time of the first group culture stage of the artificial light source of the plant tissue culture factory is 14h, the accumulated illumination time of the second group culture stage is 16h within 24h, the growth environment temperature of the tissue culture seedling (test-tube seedling) is 20-25 ℃, and the air humidity is 30-60%.
In a plant tissue culture factory, strip-shaped light sources are arranged on each layer of tissue culture rack, the distance between two adjacent artificial strip-shaped light sources is l, and the calculation formula of l is shown as (I):
in the formula (I), alpha is a light-emitting angle and the unit is DEG;
h is the height from the light source to the top end of the tissue culture bottle, and the unit is m.
In this example, α is 100 °; h is 0.15 m; l is 0.20 m.
In the plant tissue culture factory using the artificial light source as the main light source in the embodiment 2, that is, all light energy obtained in the growth process of the tissue culture seedling (test-tube seedling) is provided by the artificial light source, and does not relate to direct solar light and scattered light), the potato tissue culture seedling is cultured, and the height of the tissue culture seedling above the culture medium can grow to 6-7 cm 28-30 days after inoculation.
Example 3
The embodiment provides a light environment regulation and control method for delaying growth of potato tissue culture seedlings (test-tube seedlings), which is used in a plant tissue culture factory with an artificial light source as a main light source, namely, light energy obtained in the growth process of the tissue culture seedlings (test-tube seedlings) is provided by the artificial light source, and direct light and scattered light of the sun are not involved; when potato tissue culture seedlings (test-tube seedlings) are planted in the plant tissue culture factory, the artificial light source specifically comprises:
after 0-48 h (the first group culture stage) and 48h (the second group culture stage) of the potato tissue culture seedlings (test-tube seedlings) after subculture inoculation, the artificial light source comprises the following components in percentage by mass according to the light-mass spectrum integral: 35% of red light, 35% of blue light and 30% of green light, wherein the peak wavelength of the red light is 610-660 nm, the peak wavelength of the blue light is 430-480 nm, the peak wavelength of the green light is 540-560 nm, and the total illumination intensity of an artificial light source received by the top end of the tissue culture bottle in the first group of culture stages is 40 mu mol/(m mol)2S) direct and scattered light, the total illumination intensity of the artificial light source received by the top of the tissue culture bottle in the second tissue culture stage being 120 [ mu ] mol/(m)2S), the accumulated illumination time of the first group of culture stages of the artificial light source of the plant tissue culture factory is 14h, the accumulated illumination time of the second group of culture stages is 16h within 24h, the growth environment temperature of the tissue culture seedlings (test tube seedlings) is 19-22 ℃, and the air humidity is 30-60%.
In a plant tissue culture factory, strip-shaped light sources are arranged on each layer of tissue culture rack, the distance between two adjacent artificial strip-shaped light sources is l, and the calculation formula of l is shown as (I):
in the formula (I), alpha is a light-emitting angle and the unit is DEG;
h is the height from the light source to the top end of the tissue culture bottle, and the unit is m.
In this example, α is 120 °; h is 0.15 m; l is 0.26 m.
In the plant tissue culture factory using the artificial light source as the main light source in this embodiment 3, that is, all light energy obtained in the growth process of the tissue culture seedling (test-tube seedling) is provided by the artificial light source, and does not relate to direct solar light and scattered light), the potato tissue culture seedling is cultured, and the height of the tissue culture seedling above the culture medium 30 to 33 days after inoculation can reach 6 to 7 cm.
Example 4
The embodiment provides a light environment regulation and control method for delaying growth of potato tissue culture seedlings (test-tube seedlings), which is used in a plant tissue culture factory with an artificial light source as a main light source, namely, light energy obtained in the growth process of the tissue culture seedlings (test-tube seedlings) is provided by the artificial light source, and direct light and scattered light of the sun are not involved; when potato tissue culture seedlings (test-tube seedlings) are planted in the plant tissue culture factory, the artificial light source specifically comprises:
after 0-48 h (the first group cultivation stage) of subculture inoculation of potato tissue culture seedlings (test-tube seedlings), the artificial light source comprises the following components in percentage by mass according to the light mass spectrum integral: 30% of red light, 30% of blue light, 30% of green light and 10% of full visible light continuous spectrum; after 48h of subculture (second subculture stage), the artificial light source comprises the following light mass spectrum integral percentages: 30% of red light, 40% of blue light and 30% of green light, wherein the peak wavelength of the red light is 610-660 nm, the peak wavelength of the blue light is 430-480 nm, the peak wavelength of the green light is 540-560 nm, and the total illumination intensity of an artificial light source received by the top end of the tissue culture bottle in the first group of culture stages is 30 mu mol/(m mol)2S) direct and scattered light, the total illumination intensity of the artificial light source received by the top of the tissue culture bottle in the second tissue culture stage being 110 [ mu ] mol/(m)2S) direct and scattered light, the accumulated illumination time of the first group of culture stages in 24 hours by the artificial light source of the plant tissue culture factory is 14h, the accumulated illumination time of the second group of culture stages is 16h, and the growth ring of the tissue culture seedling (test-tube seedling)The ambient temperature is 19-22 ℃, and the air humidity is 30-60%.
In a plant tissue culture factory, strip-shaped light sources are arranged on each layer of tissue culture rack, the distance between two adjacent artificial strip-shaped light sources is l, and the calculation formula of l is shown as (I):
in the formula (I), alpha is a light-emitting angle and the unit is DEG;
h is the height from the light source to the top end of the tissue culture bottle, and the unit is m.
In this example, α is 120 °; h is 0.18 m; l is 0.32 m.
In the plant tissue culture factory using the artificial light source as the main light source in this embodiment 4, that is, all the light energy obtained during the growth process of the tissue culture seedling (test-tube seedling) is provided by the artificial light source, and does not relate to the direct light and the scattered light of the sun), the potato tissue culture seedling (test-tube seedling) can grow to 6-7 cm above the culture medium 35-38 days after inoculation.
Comparative example 1
The comparative example provides a light environment regulation and control method for delaying the growth of potato tissue culture seedlings (test-tube plantlets), which is the same as the conditions of the example 4, in a plant tissue culture factory which takes an artificial light source as a main light source, namely, all light energy obtained in the growth process of the tissue culture seedlings (test-tube plantlets) is provided by the artificial light source and does not relate to direct solar light and scattered light; when potato tissue culture seedlings (test-tube seedlings) are planted in the plant tissue culture factory, the only difference is that the artificial light sources provided for culturing the potato tissue culture seedlings (test-tube seedlings) are all continuous spectra of full visible light. The total illumination intensity and illumination time of the artificial light source received by the top end of the tissue culture bottle, the culture environment temperature and the air humidity are the same as those in the embodiment 4.
This comparative example 1 was compared with example 4 and the growth of tissue culture seedlings (test-tube seedlings) of potato was observed.
In the plant tissue culture factory in example 4 using the artificial light source as the main light source, that is, all light energy obtained during the growth of the tissue culture seedling (test-tube seedling) is provided by the artificial light source, and does not relate to direct solar light and scattered light), the potato tissue culture seedling is cultured, and the height of the tissue culture seedling above the culture medium can reach 6-7 cm after the potato tissue culture seedling (test-tube seedling) is inoculated for 35-38 days.
In the comparative example 1, in a plant tissue culture factory with an artificial light source as a main light source, that is, all light energy obtained in the growth process of the tissue culture seedlings (test-tube seedlings) is provided by the artificial light source, and does not relate to direct solar light and scattered light, the artificial light source provided for the growth of the potato tissue culture seedlings (test-tube seedlings) is a full visible light continuous spectrum, the height of the potato tissue culture seedlings (test-tube seedlings) above a culture medium can grow to 6-7 cm in 18-21 days after inoculation, and the potato tissue culture seedlings (test-tube seedlings) in the embodiment 4 can grow for 17 days more.
Comparative example 2
The comparative example provides a light environment regulation and control method for delaying the growth of potato tissue culture seedlings (test-tube plantlets), which is the same as the conditions of the example 4, in a plant tissue culture factory which takes an artificial light source as a main light source, namely, all light energy obtained in the growth process of the tissue culture seedlings (test-tube plantlets) is provided by the artificial light source and does not relate to direct solar light and scattered light; when planting potato tissue culture seedling (tube seedling) in above-mentioned plant tissue culture mill, the only difference is that the spectrum that the artifical light source that provides for cultivating potato tissue culture seedling (tube seedling) is suitable potato tissue culture seedling (tube seedling) fast growth, and in the first group banks up the stage promptly, first artifical light source includes according to the light mass spectrum integral percentage: 65% of red light, 25% of blue light and 10% of full visible light continuous spectrum; in the second culturing stage, the second artificial light source comprises the following components according to the integral percentage of the light mass spectrum: continuous spectra of 66% of red light, 16% of blue light, 13% of green light and 5% of full visible light, wherein the peak wavelength of the red light is 610-660 nm, the peak wavelength of the blue light is 430-480 nm, the peak wavelength of the green light is 540-560 nm, and the total illumination intensity and illumination time of the artificial light source received by the top end of the tissue culture bottle, the culture environment temperature and the air humidity are the same as those in the embodiment 4.
This comparative example 2 was compared with example 4 and the growth of tissue culture seedlings (test-tube seedlings) of potato was observed.
In the plant tissue culture factory in which the artificial light source is used as the main light source in the embodiment 4 (i.e., all light energy obtained in the growth process of the tissue culture seedling (test-tube seedling) is provided by the artificial light source and does not relate to direct solar light and scattered light), the potato tissue culture seedling (test-tube seedling) is cultured, and the height of the tissue culture seedling above the culture medium can reach 6-7 cm 35-38 days after inoculation.
In the comparative example 2, light energy obtained in the growth process of a plant tissue culture factory (i.e. tissue culture seedlings (test-tube seedlings)) with an artificial light source as a main light source is provided by the artificial light source, and does not relate to direct solar light and scattered light, the artificial light source provided by the growth of the potato tissue culture seedlings (test-tube seedlings) is a spectrum suitable for the rapid growth of the potato tissue culture seedlings (test-tube seedlings), the height of the potato tissue culture seedlings (test-tube seedlings) above a culture medium can grow to 6-7 cm in 14-17 days after inoculation, and the potato tissue culture seedlings (test-tube seedlings) in the example 4 can grow for 21 days more.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.